System, device and method for production of bioproduct including high density cell respirator for intensified production of adeno-associated viruses

Abstract

A cell cultivation apparatus for cultivating microorganisms and growing cells at high density is provided. The apparatus includes a membrane comprising multiple surface features on a first side of the membrane for cell placement. The surface features comprising one or more compartments within which a cell can be located. The membrane includes a material that is at least partially permeable to gas. A second side of the membrane defines a gas region. The second side of the membrane is separated from the first side of the membrane by the membrane. The apparatus further includes a media region for receiving media. The compartments are configured to at least partially reduce media flow shear forces on one or more cells in the compartments. The surface features may be ridges, protrusions, fins, wells, and/or posts.

What is claimed is: 
     
         1 . A cell cultivation apparatus comprising:
 a membrane comprising an array of fins running substantially parallel to each other, each fin of the array of fins having a substantially constant cross-section along a length of the fin, the substantially parallel fins defining grooves between adjacent fins, at least one groove on a first side of the membrane providing a compartment for cell placement;   the membrane comprising a material that is at least partially permeable to gas;   a second side of the membrane defining a gas region, the second side of the membrane being separated from the first side of the membrane by the membrane, a gas capable of passing through the membrane; and   a media region on the first side of the membrane configured for receiving media including one or more cells depositable in the compartment.   
     
     
         2 . The apparatus of  claim 1 , wherein the second side is placed on a mesh, cloth, or other open pore material to allow gas from an exterior environment to exchange with the membrane. 
     
     
         3 . The apparatus of  claim 1 , wherein the second side comprises one or more variations in its geometry that permits gas to at least partially pass below the second side when the second side is placed on a flat surface. 
     
     
         4 . The apparatus of  claim 3 , wherein the variations in geometry comprise pillars, channels, grooves, bumps, protrusions, or legs. 
     
     
         5 . The apparatus of  claim 3 , wherein the variations in geometry comprise one or more spacing pillars. 
     
     
         6 . The apparatus of  claim 1 , further comprising a top surface, the top surface comprising a sealed membrane. 
     
     
         7 . The apparatus of  claim 6 , wherein the top surface comprises a silicone based membrane. 
     
     
         8 . The apparatus of  claim 6 , further comprising a sealable port for transferring media to or from the apparatus. 
     
     
         9 . The apparatus of  claim 1 , wherein at least one of the fins has a height between about 400 μm and about 1000 μm, a width between about 300 μm and about 500 μm, and a pitch between about 200 μm and about 500 μm. 
     
     
         10 . The apparatus of  claim 9 , wherein the at least one of the fins has a height between about 700 μm and about 1000 μm. 
     
     
         11 . The apparatus of  claim 10 , wherein the at least one of the fins has a height of about 700 μm. 
     
     
         12 . The apparatus of  claim 1 , wherein at least one of the fins has a ratio of height to width to pitch of about 4-10 to about 3-5 to about 2-5. 
     
     
         13 . The apparatus of  claim 12 , wherein the ratio of height to width to pitch is about 7-10 to about 3-5 to about 2-5. 
     
     
         14 . The apparatus of  claim 13 , wherein the ratio of height to width to pitch is about 7 to about 3-5 to about 2-5. 
     
     
         15 . The apparatus of  claim 1 , wherein the grooves are an array of grooves with a substantially constant spacing along a length of the grooves between neighboring grooves. 
     
     
         16 . The apparatus of  claim 1 , wherein the fins and grooves are configured to be not parallel to a media flow direction during cell production. 
     
     
         17 . The apparatus of  claim 1 , further comprising:
 a media inlet configured to introduce liquid media into the media region creating a medial flow that is not parallel to the grooves.   
     
     
         18 . The apparatus of  claim 1 , wherein the fins and grooves are configured to be substantially perpendicular to a media flow direction during cell production. 
     
     
         19 . The apparatus of  claim 1 , further comprising:
 a media inlet configured to introduce liquid media into the media region creating a media flow that is substantially perpendicular to the grooves.   
     
     
         20 . The apparatus of  claim 1 , wherein the fins and grooves are configured to be substantially aligned with a media flow direction during cell harvesting. 
     
     
         21 . The apparatus of  claim 1 , further comprising:
 a media inlet configured to introduce liquid media into the media region creating a medial flow that is substantially aligned with the grooves.   
     
     
         22 . The apparatus of  claim 1 , wherein each fin of the fins has a top that is larger than a base such that an opening at the top of each groove is narrower than at least some points closer to a bottom of each groove. 
     
     
         23 . The apparatus of  claim 1 , wherein the membrane is configured to expand or contract in response to changes in pressure or flow associated with at least one of the gas and the liquid media in order to facilitate efficient cultivation or harvesting of cells located in the grooves in the membrane. 
     
     
         24 . A petri dish comprising:
 the apparatus of  claim 1 ; and   a sidewall forming a continuous and substantially vertical wall around a perimeter of the membrane.   
     
     
         25 . A single well plate platform comprising:
 a well; and   the apparatus of  claim 1  forming a bottom surface of the well.   
     
     
         26 . A multi-well plate comprising:
 multiple wells; and   the apparatus of  claim 1  forming a bottom surface in each of the multiple wells.   
     
     
         27 . A bioreactor comprising:
 a gas inlet;   a gas outlet;   a gas path connected between the gas inlet and the gas outlet;   a media inlet;   a media outlet;   a media path connected between the media inlet and the media outlet; and   the apparatus of  claim 1  separating the gas path and the media path.   
     
     
         28 . The apparatus of  claim 1 , wherein the fins protect one or more cells in the grooves from media flow shear forces generated by media delivery. 
     
     
         29 . The apparatus of  claim 1 , wherein the fins form cell niche regions below a top surface of the fins. 
     
     
         30 . The apparatus of  claim 1 , wherein the fins are configured to constrain a size of cell aggregates. 
     
     
         31 . The apparatus of  claim 1 , wherein a mechanical stretching of the membrane widens at least one groove of the grooves in order to facilitate a release of one or more cells from the at least one groove.